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Method for colourimetric identifying and detecting DNA molecule

A DNA molecule and colorimetric technology, applied in the field of detection of DNA molecules, to achieve the effect of quantitative detection, broad application prospects, and simple operation

Inactive Publication Date: 2006-06-14
NORTHEAST NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no report on the development of color-changing molecular ordered assemblies with DNA molecular recognition and detection functions by using polymerized diacetylene lipid films

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Probe DNA1: 5'-TCTCAACTCGTATTTTTT-cholesteryl-3'

[0032] Probe DNA2: 5'-cholesteryl-TTTTTTCGCATTCAGGAT-3'

[0033] cDNA: 5'-TACGAGTTGAGAATCCTGAATGCG-3'

[0034] mDNA: 5'-GCGTAACTCCTAAGAGTTGAGCTA-3'

[0035] mDNA1: 5'-TACCTCTACAGAATCCTGAATGCG-3'

[0036] mDNA2: 5'-TAGCTGTTGAGAATCCTGAATGCG-3'

[0037] mDNA3: 5'-TAGGAGTTGAGAATCCTGAATGCG-3'

[0038] Preparation of vesicle 1: (1), probe DNA1 (mole percent 10%), 10,12-tricosyldiynoic acid or 10,12-pentacosyldiynoic acid (mole percent 70%) %) and a chloroform solution of phospholipids (20% by mole%) are mixed, and the organic solvent is evaporated by rotary evaporation; (2), adding distilled water or buffer solution (pH=7.4), so that the molar concentration of diacetylene lipid is 1mM; ( 3), ultrasonically disperse at 60°C for 55min; (4), store the dispersion obtained in (3) overnight at 4°C; Irradiate and polymerize for 10 minutes to obtain blue polymerized diacetylene vesicles 1 functionalized with probe DNA molecules...

Embodiment 2

[0043] Probe DNA1: 5'-TCTCAACTCCGGTTTTTT-cholesteryl-3'

[0044] Probe DNA2: 5'-cholesteryl-TTTTTTTAAGATTCAGGAT-3'

[0045] cDNA: 5'-CCGGAGTTGAGAATCCTGAATTTC-3'

[0046] mDNA: 5'-GCGTAACTCCTAAGAGTTGAGCTA-3'

[0047] mDNA1: 5'-CACCAGTAGAGAATCCTGAATTTC-3'

[0048] mDNA2: 5'-CCTTAGTTGAGAATCCTGAATTTC-3'

[0049] mDNA3: 5'-CTGGAGTTGAGAATCCTGAATTTC-3'

[0050] Preparation of vesicle 1: (1), probe DNA1 (5% by mole), 5,7-tricosyldiynoic acid or 5,7-pentacosyldiynoic acid (50% by mole %) and a chloroform solution of phospholipids (45% by mole%) were mixed, and the organic solvent was evaporated by rotary evaporation; (2), adding distilled water or a buffer solution (pH=9), so that the molar concentration of the diacetylene lipid was 1 mM; ( 3), ultrasonically disperse at 50°C for 30min; (4), store the dispersion obtained in (3) overnight at 4°C; Irradiate and polymerize for 10 minutes to obtain blue polymerized diacetylene vesicles 1 functionalized with probe DNA molecules 1 .

...

Embodiment 3

[0055] Probe DNA1: 5'-GGTTAACTCGTATTTTTT-cholesteryl-3'

[0056] Probe DNA2: 5'-cholesteryl-TTTTTTCGCATTCAGGCC-3'

[0057] cDNA: 5'-TACGAGTTAACCGGCCTGAATGCG-3'

[0058] mDNA: 5'-GCGTAACTCCTAAGAGTTGAGCTA-3'

[0059] mDNA1: 5'-TAGATGTCCACCGGCCTGAATGCG-3'

[0060] mDNA2: 5'-TACGAGTTAACCGGCCTGAATGCG-3'

[0061] mDNA3: 5'-TACCAGTTAACCGGCCTGAATGCG-3'

[0062] Preparation of vesicle 1: (1), probe DNA1 (2% by mole), 10,12-tricosyldiynoic acid or 10,12-pentacosyldiynoic acid (98% by mole %) chloroform solutions were mixed, and the organic solvent was evaporated by rotary evaporation; (2), distilled water or buffer solution (pH=5) was added, so that the molar concentration of the diacetylene lipid was 1 mM; (3), under the condition of 40° C. Ultrasonic dispersion for 20 minutes; (4), the dispersion obtained by (3) was stored overnight at 4°C; (5) the dispersion of (4) was irradiated with 256m ultraviolet light for 10 minutes to obtain probe DNA molecules 1 functionalized blue poly...

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Abstract

The invention discloses a method for colorimetric recognition and detection of DNA molecules, which belongs to the field of biomolecular detection. In the present invention, the functionalized polymerized diacetylene vesicles obtained by using two kinds of probe DNA respectively play the dual roles of color-changing indicator group and color-changing enhancement in detection. The invention is simple and fast in operation; it can realize the rapid qualitative detection of DNA molecules without the aid of instruments (colorimetric observation with naked eyes); it can realize the new method of rapid quantitative detection of DNA molecules by means of ordinary ultraviolet-visible spectrometers.

Description

technical field [0001] The invention belongs to biomolecule detection methods, in particular to a method for detecting DNA molecules. Background technique [0002] Commonly used methods for identifying and detecting DNA molecules include radioactive labeling, fluorescent labeling, enzyme labeling, and chemiluminescent methods. Most of these methods involve some expensive detection instruments and equipment, and there is also the possibility of radioactive contamination for radioactive labels. Therefore, the development of radiation-free background, simple operation, rapid, economical, simple equipment or no instrument [0003] New methods for the identification and detection of DNA molecules in organotypic devices have important practical significance. [0004] In recent years, the research on molecular ordered assemblies has developed rapidly, which provides a favorable tool for the use of bionic technology to study new methods with biomolecular recognition. Among them, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 马占芳王春刚
Owner NORTHEAST NORMAL UNIVERSITY